Detection and Localization of Anti and Pro-apoptotic mRNA Genes in Human Colorectal Cancer Using in situ RT-PCR
- 1 University Industry Selangor, Malaysia
- 2 University Putra Malaysia, Malaysia
- 3 Cyberjaya University College of Medical Sciences, Malaysia
Abstract
Problem statement: Recent studies using conventional Polymerase Chain Reaction have shown that anti-apoptotic (Cyclooxygenase-2, COX-2 and Nuclear Factor-κB, NF-κB) and proapoptotic mRNA (Bax and Bad) are involved in the tumorigenesis of colorectal cancer. The aim of this study was to localize the expression of anti and pro-apoptotic mRNA genes using Reverse Transcription in situ Polymerase Chain Reaction (RT in situ PCR) and immunodetection technique in the early stage of human colorectal adenocarcinoma. Approach: Reverse Transcription in situ Polymerase Chain Reaction (RT in situ PCR) and immunodetection technique was applied throughout of this studies. 20 paraffin-embedded tissue blocks of human colorectal adenocarcinoma samples was used compared to controls. Results: Morphologically, the glands and crypts were well-differentiated, enlarged and irregular with active secretion of mucin. COX-2, NF-κB, Bax and Bad mRNA were expressed in both normal and human colorectal cancer tissues. All mRNA genes were expressed in the cytoplasm and nuclei. However, COX-2 and NF-κB mRNA genes were highly expressed with higher intensity of brown staining compared to Bax and Bad at tubular epithelium cells. Conclusion: This study demonstrated that by using RT in situ PCR, COX-2 and NF-κB mRNA genes were shown to be involved in the development of human colorectal cancer.
DOI: https://doi.org/10.3844/ajbbsp.2010.164.171
Copyright: © 2010 Mohd. Nazil Salleh, Boo Siaw Shi, Wan Khairuzzaman Wan Ramli, Azlina Zahari, Norashikin Shamsudin and Thuaibah Hashim. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
- 3,934 Views
- 3,052 Downloads
- 0 Citations
Download
Keywords
- Cyclooxygenase-2 (COX-2)
- Nuclear Factor-κB (NF-κB)
- heterodimeric partner for Bcl- XL and Bcl-2 (Bad)
- bcl-2 associated X protein (Bax)
- in situ Reverse Transcription Polymerase Chain Reaction (in situ RT-PCR)